NMDA NR2B antagonists for treatment

ABSTRACT

The invention provides new methods for treating certain disorders resulting from neurodegeneration and for treating depression which comprise administration of NR2B subunit selective NMDA antagonists. The disorders that can be treating by the invention include hearing loss, vision loss, neurodegeneration caused by epileptic seizures, neurotoxin poisoning, Restless Leg Syndrome, multi-system atrophy, non-vascular headache, and depression.

This application claims priority under 35 U.S.C. 119(e) of U.S.Provisional Application No. 60/237,770, filed Oct. 2, 2000.

FIELD OF THE INVENTION

This invention relates to the treatment of neurological disorders. Thisinvention also relates to the treatment of depression. Moreparticularly, this invention relates to treatment of hearing loss,vision loss, neurodegeneration caused by epileptic seizures, neurotoxinpoisoning, Restless Leg Syndrome, multi-system atrophy, non-vascularheadache, and depression comprising administering anN-methyl-D-aspartate (NMDA) NR2B subtype receptor antagonist.

BACKGROUND OF THE INVENTION

NMDA Receptors and NMDA Receptor Subunits

Glutamate and aspartate play dual roles in the central nervous system asessential amino acids and as the principal excitatory neurotransmitters(hereinafter referred to as excitatory amino acids or EAAS). There areat least four classes of EAA receptors: NMDA, AMPA(2-amino-3-(methyl-3-hydroxyisoxazol-4-yl)propanoic acid), kainate andmetabotropic receptors. These EAA receptors mediate a wide range ofsignaling events that impact all physiological brain functions. Forexample, it has been reported that NMDA receptor antagonists produce ananalgesic effect under certain conditions (Wong, C. S., Cherng, C. H.and Ho, S. T., Clinical Applications of Excitatory Amino AcidAntagonists in Pain Management Acta Anaesthesiologica. Sinica; 33,227-232 (1995)).

The NMDA receptor is an ion channel permeable to Na⁺ and Ca²⁺. Thereceptor is gated by synaptically released glutamate in the presence ofco-agonist glycine and concomitant depolarization (Mayer, M. L. andWestbrook, G. L., The Physiology of Excitatory Amino Acids in theVertebrate Nervous System, Progress in Neurobiology, 28, 197-276(1987)). Thus, NMDA receptor activity may be attenuated by blockade, forexample, of 1) the glutamate binding site, 2) the glycine co-agonistbinding site, or 3) the site of the ion channel.

The NMDA receptor is composed of multiple protein subunits (Seeburg, P.H., The Molecular Biology of Mammalian Glutamate Receptor Channels,Trends in Neurosci., 16, 359-365 (1993)). The protein subunits fall intotwo categories: NR2 and NR1. The NR2 subunits contain glutamate bindingsites, whereas the NR1 subunits contain the glycine binding sites. Fivesubunits have been cloned to date, namely NR1 and NR2A, NR2B, NR2C andNR2D. Expression studies indicate the functional receptor is composed ofat least one NR1 site and one or more of the NR2 sites. Thus, differentsubtypes of NMDA receptors can be categorized based on their particularNR2 subunit composition. For example, in the adult mammalian brain, theNR1 and NR2A subunits are widely expressed, forming a subtype of NMDAreceptor comprising an NR2A subunit. In contrast, NR2B subunitexpression is mostly localized in forebrain regions including cortex,hippocampus and striatum; the NR2C subunit is expressed in thecerebellum; and the NR2D subunit is restricted to the midbrain region.NMDA receptor subtypes of corresponding composition can accordinglyrespectively be found in forebrain, cerebellum, and midbrain.

Compounds that inhibit NMDA receptor activity by interacting at theglutamate, glycine, or receptor-associated ion channel as describedabove have little (<10-fold) selectivity across the different NMDAreceptor subtypes. That is, such compounds inhibit NMDA receptors withpotencies within a 10-fold range regardless of the subunit combination.However, the subunit composition of the NMDA receptor can confer uniquephysiology with regard to conductance, kinetics, and affinity forcertain agonists. For example, the subunit composition of an NMDAreceptor has significant effects on its sensitivity to a group ofallosteric modulators which include protons, polyamines, Zn²⁺, andoxidizing/reducing agents (Chenard, B. L. and Menniti, F. S.,Antagonists Selective for NMDA Receptors Containing the NR2B Subunit,Current Pharmaceutical Design, 1999, 5:381-404)). Receptors comprisingthe NR2B subunit possess a unique site to which compounds may bind,resulting in specific inhibition this subtype of NMDA receptor ascompared to NMDA receptors that do not comprise an NR2B subunit (Ibid).This unique site is distinct from the glutamate binding site on the NR2Bsubunit.

Antagonizing NMDA receptors at the NR2B subunit specific binding sitecan be used to substantially avoid side effects that have been noted attherapeutic drug levels with other non-specific NMDA receptorantagonists. Both glutamate competitive antagonists and channel blockingagents cause cardiovascular effects and psychotic symptoms in man(Chenard and Menniti, supra). In rodents, these types of compounds alsocause locomotor hyperactivity and a paradoxical neuronalhyperexcitability manifest as neuronal vacuolization in cingulate andretrosplenial cortices (Id.). Antagonists at the glycine co-agonist sitecause less locomoter activation and do not cause neuronal vacuolizationat neuroprotective doses in rodents, however physicochemical problems(for example, problems relating to solubility, brain penetration andprotein binding) associated with the quinoxalinedione nucleus typical ofsuch compounds have hindered efforts to bring this class of moleculesforward in the clinic (Id). NMDA receptor antagonists selective for theNR2B subunit offer a means of inhibition without the side effects andpsychochemical difficulties described above.

NR2B Subunit Selective NMDA Receptor Antagonists

Compounds that inhibit NMDA receptors comprising an NR2B subunit byspecific binding to the NR2B subunit have been demonstrated bymeasurement of inhibition of NMDA-induced current in Xenopus Oocytescotransfected with the genes expressing the NR1 and NR2B subunits(Chenard and Menniti, supra). Specificity for NR2B can be confirmed byobserving reduced inhibition of the NMDA-induced current in XenopusOocytes cotransfected with an NR1 subunit and an NR2 subunit other thanNR2B.

A number of compounds have been found to act as antagonists that targetthe NR2B subunits of NMDA receptors that contain them. The firstcompound identified to display significant affinity for the NR2B subunitwas ifenprodil. Ifenprodil is both more potent and efficacious forblockade of ion current through NMDA receptors comprised of NR1/NR2Bsubunits compared to NR1/NR2A, NR2C, or NR2D subunits.

For example, ifenprodil and related compounds have been demonstrated inanimal models of pain perception to produce significant analgesicactivity (Bernardi, M., Bertolini, A., Szczawinska, K. And Genedani, S.,Blockade of the Polyamine Site of NMDA Receptors ProducesAntinociception and Enhances the Effect of Morphine, in Mice, EuropeanJournal of Pharmacology, 298, 51-55, (1996); Taniguchi, K., Shinjo, K.,Mizutani, M., Shimada, K., Ishikawa, T., Menniti, F. S. and Nagahisa, A,Antinociceptive Activity of CP-101,606, an NMDA Receptor NR2B SubunitAntagonist, British Journal of Pharmacology, 122, 809-812 (1997)).

U.S. Pat. No. 5,710,168 (issued Jan. 20, 1998) claims the use of certaincompounds of formula I, infra, having NR2B subunit selectivity fortreating a disease or condition which is susceptible to treatment byblocking of NMDA receptor sites, including traumatic brain injury,spinal cord trauma, pain, psychotic conditions, drug addiction,migraine, hypoglycemia, anxiolytic conditions, urinary incontinence, andischemic events arising from CNS surgery, open heart surgery or anyprocedure during which the function of the cardiovascular system iscompromised.

U.S. Ser. No. 09/397,891, filed Sep. 17, 1999, pertains to a method oftreating acute, chronic and/or neuropathic pain comprising administeringan NR2B selective NMDA receptor antagonist, for example a compound offormula I, infra.

U.S. Pat. No. 5,710,168 and U.S. Ser. No. 09/397,891 are bothincorporated by reference herein in their entireties.

SUMMARY OF THE INVENTION

The present invention provides a method for treating sensorineuralhearing loss in a mammal, which method comprises administering to themammal an amount of an NR2B subunit selective NMDA antagonist, whichamount is effective in treating sensorineural hearing loss.

This invention also provides a method for treating neurological damagecaused by epileptic seizures in a mammal, which method comprisesadministering to the mammal an amount of an NR2B subunit selective NMDAantagonist, which amount is effective in inhibiting neurological damage.

This invention further provides a method for treating neurologicaldamage caused by neurotoxin poisoning in a mammal, which methodcomprises administering to the mammal an amount of an NR2B subunitselective NMDA antagonist, which amount is effective in inhibitingneurological damage.

This invention further provides a method for treating vision loss causedby neurodegeneration of the visual pathway in a mammal, which methodcomprises administering to the mammal an amount of an NR2B subunitselective NMDA antagonist, which amount is effective in treating visionloss caused by neurodegeneration of the visual pathway.

This invention also provides a method of treating Restless Leg Syndromein a mammal, which method comprises administering to the mammal anamount of an NR2B subunit selective NMDA antagonist, which amount iseffective in treating Restless Leg Syndrome.

This invention also provides a method of treating multi-system atrophyin a mammal, which method comprises administering to the mammal anamount of an NR2B subunit selective NMDA antagonist, which amount iseffective in treating multi-system atrophy.

This invention also provides a method of treating non-vascular headachein a mammal, which method comprises administering to the mammal anamount of an NR2B subunit selective NMDA antagonist, which amount iseffective in treating non-vascular headache.

This invention also provides a method of treating depression in amammal, which method comprises administering to the mammal an amount ofan NR2B subunit selective NMDA antagonist, which amount is effective intreating depression.

In one embodiment, the NR2B subunit selective NMDA antagonist in each ofthe preceding methods is a compound of formula I

or a pharmaceutically acceptable acid addition salt thereof or anenantiomer thereof, wherein:

-   -   (a) R² and R⁵ are taken separately and R¹, R², R³ and R⁴ are        each independently hydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷        and R⁵ is methyl or ethyl; or    -   (b) R² and R⁵ are, taken together,    -    thereby forming a chroman-4-ol ring, and R¹, R³ and R⁴ are each        independently hydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷;    -   R⁶ is    -   R⁷ is methyl, ethyl, isopropyl or n-propyl;    -   R⁸ is phenyl optionally substituted with up to three        substituents independently selected from the group consisting of        (C₁-C₆) alkyl, halo and CF₃;    -   X is O, S or (CH₂)_(n); and    -   n is 0, 1, 2, or 3.

In another embodiment of each of the preceding methods, the NR2B subunitselective NMDA antagonist is:

-   -   (+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;

-   (1S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;

-   (3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-chroman-4,7-diol;    or

-   (1R*,2R*)-1-(4-hydroxy-3-methylphenyl    )-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol; or    -   an enantiomer of one of the aforementioned compounds; or    -   a pharmaceutically acceptable acid addition salt of one of the        aforementioned compounds or one of their enantiomers.

DETAILED DESCRIPTION OF THE INVENTION

“Mammal” as used herein refers to any mammal, including humans.

The phrase “sensorineural hearing loss” refers to hearing loss caused byloss of neurons. Such hearing loss can be, for example, genetic inorigin. Another example of sensorineural hearing loss isantibiotic-induced, such as aminoglycoside-induced, hearing loss.Sensorineural hearing loss can also be induced by excessive sound.

“Neurotoxin poisoning” refers to poisoning caused by a neurotoxin. Aneurotoxin is any chemical or substance that can cause neural death andthus neurological damage. An example of a neurotoxin is alcohol, which,when abused by a pregnant female, can result in alcohol poisoning andneurological damage known as Fetal Alcohol Syndrome in a newborn. Otherexamples of neurotoxins include, but are not limited to, kainic acid,domoic acid, and acromelic acid; certain pesticides, such as DDT;certain insecticides, such as organophosphates; volatile organicsolvents such as hexacarbons (e.g. toluene); heavy metals (e.g. lead,mercury, arsenic, and phosphorous); aluminum; certain chemicals used asweapons, such as Agent Orange and Nerve Gas; and neurotoxicantineoplastic agents.

“Neurodegeneration of the visual pathway” refers to neural cell deathoccurring in neurons involved in vision, for example neurons in theoccipital lobe, optic nerve, and retina. Neurodegeneration of the visualpathway can be caused, for example, by a stroke in the visual pathway,for example a retinal stroke. Stoke can also occur in the optic nerve orthe occipital lobe. Neurodegeneration of the visual pathway can also becaused by neurodegenerative diseases, for example macular degeneration.Neurodegeneration of the visual pathway can also be caused by diseasesthat are not necessarily considered neurodegenerative, such as glaucoma,which can cause retinal degeneration.

“Non-vascular headache” generally refers to headaches other thanmigraines. Examples of non-vascular headaches include, but are notlimited to, stress headaches and sinus headaches.

The phrase “neurological damage” refers herein to neuron cell death.

The terms “treatment”, “treating”, and the like, refer to reversing,alleviating, or inhibiting the progress of the disease or condition towhich such term applies, or one or more symptoms of such disease orcondition. As used herein, these terms also encompass, depending on thecondition of the patient, preventing the onset of a disease orcondition, or of symptoms associated with a disease or condition. Suchprevention also includes reducing the severity of a disease or conditionor symptoms associated therewith prior to affliction with said diseaseor condition. Thus, “treatment” encompasses administration of theantagonist to a subject that is not at the time of administrationafflicted with the disease or condition, and “treatment” can includepreventing the recurrence of a disease or condition or of symptomsassociated therewith. Conditions wherein a patient who is not at thetime of examination afflicted with a disease or condition but couldbenefit from treatment according to a method described herein can berecognized by a healthcare professional, such as a medical doctor, ofordinary skill.

NR2B subunit selective NMDA antagonists that can be used in the methodsof the present invention include compounds of formula I

and pharmaceutically acceptable acid addition salt thereof, wherein:

-   -   (a) R² and R⁵ are taken separately and R¹, R², R³ and R⁴ are        each independently hydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷        and R⁵ is methyl or ethyl; or    -   (b) R² and R⁵ are, taken together,    -    thereby forming a chroman-4-ol ring, and R¹, R³ and R⁴ are each        independently hydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷;    -   R⁶ is    -   R⁷ is methyl, ethyl, isopropyl or n-propyl;    -   R⁸ is phenyl optionally substituted with up to three        substituents independently selected from the group consisting of        (C₁-C₆) alkyl, halo and CF₃;    -   X is O, S or (CH₂)_(n); and    -   n is 0, 1, 2, or 3.

Specific compounds of formula I that can be used are:

-   (+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;-   (1S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;-   (3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-chroman-4,7-diol;    -   pharmaceutically-acceptable salts of the above compounds; and-   (1R*,2R*)-1-(4-hydroxy-3-methylphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol;    -   and enantiomers of any of the aforementioned compounds;    -   and pharmaceutically acceptable acid addition salts of any of        the aforementioned compounds and of any of their enantiomers.

The compounds of formula I can be prepared as follows. The compounds offormula I wherein R² and R⁵ are taken together forming a chroman-4-olring, and R¹, R³, and R⁴ are hydrogen, can be prepared by one or more ofthe synthetic methods described and referred to in U.S. Pat. No.5,356,905 (incorporated herein by reference, supra). The compounds offormula I wherein R² and R⁵ are taken separately, and R¹, R², R³ and R⁴are hydrogen can be prepared by one or more of the synthetic methodsdescribed and referred to in U.S. Pat. Nos. 5,185,343; 5,272,160; and5,338,754; all of which are incorporated herein by reference in theirentireties. The compounds of formula I can also be prepared by one ormore of the synthetic methods described and referred to in U.S. patentapplication Ser. No. 08/292,651; U.S. Pat. No. 5,744,483 (issued Apr.28, 1998) and U.S. Pat. No. 6,008,233 (issued Dec. 28, 1999); PCTInternational Application No. PCT/IB95/00398 which designates the UnitedStates (filed May 26, 1995) (corresponding to WO 96/37222); and PCTInternational Application No. PCT/IB95/00380 which designates the UnitedStates (filed May 18, 1995) (corresponding to WO 96/06081). These UnitedStates patents and PCT International Applications, and the United Statespatent application, are also all incorporated by reference herein intheir entireties.

A preferred compound,(1S,2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-1-propanol((1S,2S) free base), and its tartrate salt, can be prepared as describedin U.S. Pat. No. 5,272,160, referred to above. The resolution of racemic1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-1-propanol toform the (1S,2S) free base and the corresponding (1R,2R) enantiomer canbe carried out as described in U.S. Pat. No. 6,008,233 (issued Dec. 28,1999), referred to above, and as exemplified in Example 1 below.

The anhydrous mesylate of the (1S,2S) free base can be prepared asdescribed in U.S. Pat. No. 5,272,160, referred to above. The anhydrousmesylate of the (1S,2S) free base, when equilibrated in an 81% relativehumidity environment, will convert to the mesylate salt trihydrate ofthe (1S,2S) enantiomer.

The mesylate salt trihydrate of(1S,2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-1-propanolcan be prepared from the (1S,2S) free base as described in the U.S. Pat.No. 6,008,233, entitled“(1S,2S)-1-(4-Hydroxyphenyl)-2-(4-Hydroxy-4-Phenylpiperidin-1-yl)-1-PropanolMethanesulfonate Trihydrate”, referred to above and incorporated hereinby reference in its entirety. In this method, (1S,2S) free base isdissolved in water at 30° C. To this solution is added at least 1equivalent of methane sulfonic acid and the resulting mixture is warmedto 60-65° C. The warm solution can be filtered to render it particulatefree. The solution is concentrated to approximately 40% of the initialvolume, cooled below 10° C., isolated by filtration and dried to a watercontent (measured Karl Fischer titration) of approximately 11.3%. Theresulting crystalline mesylate salt trihydrate can be further purifiedby recrystallization.

Another preferred compound,(3R,4S)-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4,7-diol((3R,4S) chromanol), can be prepared as described in U.S. Pat. Nos.5,356,905, 5,744,483 (issued Apr. 28, 1998), and U.S. provisional patentapplication entitled “Process For The Resolution Of Cis-Racemic7-Benzyloxy-3-[4-(4-Fluorophenyl)-4-Hydroxy-Piperidin-1-yl]-Chroman-4-olDibenzoyl-D-Tartrate”, all three of which are referred to above. Thestarting materials and reagents required for the synthesis of the(3R,4S) chromanol are readily available, either commercially, accordingto synthetic methods disclosed in the literature, or by syntheticmethods exemplified in the description provided below.

The (3R,4S) chromanol can be prepared by fractional crystallization ofthe L-proline ester of racemiccis-7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4-ol,as described in U.S. Pat. No. 5,744,483, referred to above. In apreferred method, the resolution method described in U.S. provisionalpatent application entitled “Process For The Resolution Of Cis-Racemic7-Benzyloxy-3-[4-(4-Fluorophenyl)-4-Hydroxy-Piperidin-1-yl]-Chroman-4-olDibenzoyl-D-Tartrate”, referred to above, and as exemplified in Example3. In this method, the parent chromanol is prepared by dissolvingracemiccis-7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4-olwith an equal molar amount of dibenzoyl-D-tartaric acid in boilingaqueous ethanol. Racemiccis-7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4-olis prepared as described in U.S. patent application Ser. No. 08/189,479,referred to above. The concentration of aqueous ethanol is not criticaland may be varied between 75% and 95% ethanol (ETOH). A concentration of9:1/ETOH:H₂O has been found to be effective and is preferred. Asufficient amount of the aqueous ethanol solvent to dissolve the racemiccompound is required. This amount has been found to be about 17 ml pergram of racemic compound.

Upon stirring while heating under reflux, the racemic compound dissolvesto form a hazy solution which is allowed to cool with stirring whereuponthe (+) isomer,(3R,4S)-7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-yl]-chroman-4-oldibenzoyl-D-tartrate, precipitates and may be collected by filtrationand washed with aqueous ethanol. This is the tartrate salt of the(3R,4S) chromanol. The lactate and mandelate salts of the (3R,4S)chromanol are prepared in an analogous manner. This initial product isof about 90% optical purity. If a higher purity is desired, the productmay be heated again with aqueous ethanol, cooled and the productcollected and washed. Two such treatments were found to yield the (+)isomer of 99.4% optical purity in an overall yield of 74%. This methodavoids a reduction step with lithium aluminum hydride and is thereforepreferable for bulk operations. This method also can produce asignificantly higher yield of the desired product.

The above described (+) isomer can be converted to(3R,4S)-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4,7-diolby standard procedures. For example, treatment with dilute base can beused to free the piperidinyl base and subsequent hydrogeneration removesthe 7-benzyl group to yield the (3R,4S) chromanol.

NR2B subunit selective NMDA receptor antagonists useful in the practiceof the invention may also be used in the form of a pharmaceuticallyacceptable salt. The expression “pharmaceutically-acceptable acidaddition salts” is intended to include but not be limited to such saltsas the hydrochloride, hydrobromide, sulfate, hydrogen sulfate,phosphate, hydrogen phosphate, dihydrogenphosphate, acetate, succinate,citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) andp-toluenesulfonate (tosylate) salts. The acid addition salts of thecompounds of the present invention are readily prepared by reacting thebase forms with the appropriate acid. When the salt is of a monobasicacid (e.g., the hydrochloride, the hydrobromide, the p-toluenesulfonate,the acetate), the hydrogen form of a dibasic acid (e.g., the hydrogensulfate, the succinate) or the dihydrogen form of a tribasic acid (e.g.,the dihydrogen phosphate, the citrate), at least one molar equivalentand usually a molar excess of the acid is employed. However when suchsalts as the sulfate, the hemisuccinate, the hydrogen phosphate or thephosphate are desired, the appropriate and exact chemical equivalents ofacid will generally be used. The free base and the acid are usuallycombined in a co-solvent from which the desired salt precipitates, orcan be otherwise isolated by concentration and/or addition of anon-solvent.

Any other compound that is an NR2B subunit selective NMDA receptorantagonist, including its pharmaceutically acceptable salts, can be usedin the methods of this invention. NMDA receptor antagonists having NR2Bsubunit selectivity that may be used according to the present inventionare, for example, described in U.S. Pat. Nos. 6,046,213; 5,185,343;5,272,160, 5,338,754; and 5,356,905 (which issued, respectively, on Apr.4, 2000; Feb. 9, 1993; Dec. 21, 1993; Aug. 16, 1994; and Oct. 18, 1994);U.S. Pat. No. 6,046,213 (issued Apr. 4, 2000), U.S. Pat. No. 5,744,483(issued Apr. 28, 1998) and U.S. Pat. No. 6,008,233 (issued Dec. 28,1999); PCT International Application No. PCT/IB95/00398 (filed May 26,1995, corresponding to WO 96/37222); and PCT International ApplicationNo. PCT/IB95/00380 (filed May 18, 1995, corresponding to WO 96/06081).Other NR2B subunit selective NMDA receptor antagonists that may be usedaccording to the present invention are described in WO 97/32581(International Publication Date Sep. 12, 1997), WO 98/18793(International Publication Date May 7, 1998), WO 97/23202 (InternationalPublication Date Jul. 3, 1997), EP 0 824 098 A1 (Date of Publication,Feb. 18, 1998), EP 0846 683 A1 (Date of Publication, Jun. 10, 1998), andDE 19739331 (published Nov. 26, 1998). All of the foregoing patents andpublished patent applications are incorporated by reference herein intheir entireties.

Other compounds that are indicated to bind selectively to NR2B NMDAreceptor subunits that may be used according to the present inventionare ifenprodil, supra, eliprodil (described in U.S. Pat. No. 4,690,931(issued Sep. 1, 1987); and compounds described in WO 97/23458(International Publication Date Jul. 3, 1997), WO 97/23216(International Publication Date Jul. 3, 1997); WO 97/23215(International Publication Date Jul. 3, 1997); and WO 97/23214(International Publication Date Jul. 3, 1997). The preceding U.S. patentand PCT International Applications are incorporated by reference hereinin their entireties.

WO 97123458 discloses 2-substituted piperidine analogs. Examples ofthese analogs include N-(methoxycarbonylbutyl)-4-benzylpiperidine,4-benzylpiperidylpentamide, 4-benzylpiperidylpentahydrazide,N-(5-(4-benzylpiperidin-1-yl)valeroyl)-N′-(4-nitrobenzoyl) hydrazine,N-(4-aminobenzoyl)-N′-(5-(4benzylpiperidin-1-yl) valeroyl) hydrazine,4-benzyl-1-(3-butynyl) piperidine,1-(3-butynyl)-4-(p-chlorophenyl)-4-hydroxypiperidine,m-fluorobenzyltriphenylphosphonium bromide,1-benzyl-4-(m-fluorobenzylidene) piperidine,4-(4-benzylpiperidinyl)-4′-fluorobenzophenone and4,4′-bis(4-benzylplperidinyl) benzophenone.

WO 97/23216 discloses 4-substituded piperidine analogs. Examples ofthese analogs include 4-benzyl-1-(2-phenoxyethyl) piperidine,4-benzyl-1-(3-phenoxypropyl) piperidine,4-benzyl-1-[2-hydroxy-3-(1-naphthyloxy)propyl]piperidine,4-benzyl-1-[(2-hydroxy-4-phenyl)butyl] piperidine,4-phenoxy-1-[(4-fluorophenoxy)propyl]piperidine,4-(2-methoxyphenoxy)-1-(4-phenylbutyl) piperidine,1-(3-phenoxypropyl)-4-phenylpiperidine,1-(2-phenoxyethyl)-4-phenylpiperidine,1-(4-phenoxybutyl)-4-phenylpiperidine, and1-(4-(3-(trifluoromethyl)phenoxy)butyl)-4-phenylpiperidine hydrobromide.

WO 97/23215 discloses 2-substituted piperidine analogs. Examples ofthese analogs include2-benxylpiperidine hydrochloride,1,2-dibenzylpiperidine, hydrobromide, 2-benzyl-1-(2-phenylethyl)piperidine, hydrobromide, 2-benzyl-1-(3-phenylpropyl) piperidine,hydrobromide, 2-benzyl-1-(4-phenylbutyl) piperidine), citric acid salt,2-benzyl-1-(2-phenoxyethyl) piperidine, citric acid salt,2-benzyl-1-(3-phenoxypropyl) piperidine hydrobromide,2-benzyl-1-(4-phenoxybutyl) piperidine hydrobromide, and4-(o-(trifluoromethyl)phenoxy)-1-bromobutane.

WO 97/23214 discloses 4-substituded piperidine analogs, includinghydroxypiperidine and tetrahydropyridine analogs. Examples of theseanalogs include 1-(3-butynyl)-4-(4-chlorobenzyl) piperidine,1-(3-butynyl)-4-[(4-trifluoromethylphenyl)methyl]piperidine,4-(4-chloro)benzyl-1-[4-(3-aminophenyl)-3-butynyl]-piperidine,4-(4-trifluoromethyl)benzyl-1-[4-(3-aminophenyl)-3-butynyl]-piperidine,1-(t-butyldimethylsiloxy)-4-(3-amino-phenyl)-3-butynyl,1-tosyl-4-(3-aminophenyl)-3-butyne,1-[4-(5-(2-amino)pyridynyl)-3-butynyl]-4-hydroxy-4-(4-chlorophenyl)piperidine, 1-[4-(3-aminophenyl)-3-butynyl]-4-hydroxy-4-(4-piperidine,1-(t-butyldimethylsiloxy)-4-(3-amino-phenyl)-3-butyne,1-tosyl-4-(3-aminophenyl)-3-butyne,1-[4-(5-(2-amino)pyridynyl)-3-butynyl]-4-hydroxy-4-(4-chlorophenyl)piperidine, 1-[4-(3-aminophenyl)-3-butynyl]-4-hydroxy-4-(4-chlorophenyl)piperidine,1-[4-(3-aminophenyl)-3-butynyl]-4-(4-chlorophenyl)-1,2,5,6-tetrahydropyridine,and1-[4-(5-(2-amino)pyridynyl)-3-butynyl]-4-(4-chlorophenyl)-1,2,5,6-tetrahydropyridine.

Compounds that selectively antagonize NMDA receptors comprising an NR2Bsubunit by specifically binding to the NR2B subunit can be determined byscreening compounds for inhibition of NMDA-induced current inrecombinant Xenopus Oocytes cotransfected with the NR1A subunit and theNR2B subunit (see, e.g., Monyer, et al., Science, 1992, 256:1217-1221).A compound's activity in inhibiting current in the recombinant cellscomprising the NR2B subunit can be compared to its activity inhibitingNMDA-induced current in recombinant Xenopus Oocytes expressing the NR1subunit and NR2A, NR2C, and NR2D subunits. (See, Chenard and Menniti,supra).

One general method that can also generally predict whether or not acompound has NR2B subunit selectivity, for purposes of the presentinvention, is a standard competitive binding assay using [³H]radiolabeled racemic CP-101,606 (which contains[³H](+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;see, for example, U.S. Pat. No. 6,046,213). If a compound has an IC₅₀ ofless than about 5 μM for inhibition of racemic [³H] CP-101,606 bindingto the NR2B subunit, than the compound has NR2B subunit selectivity forpurposes of the present invention. An example of such an assay is asfollows:

Example of NR2B subunit binding assay. Selectivity of compounds for theNR2B-subunit containing NMDA receptor can be defined as an affinity forthe racemic [³H] CP-101,606 binding site in forebrain of rats, asdescribed in Chenard and Menniti, supra. This affinity is assessed in aradioligand binding assay as described below. Selective compounds arepreferably those which displace specific binding of racemic[³H]CP-101,606 from rat forebrain membranes with an IC₅₀ of about≦5 μM.

The binding of racemic[³H](+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanolto rat forebrain membranes is measured as described by Menniti et al.(CP-101,606, a potent neuroprotectant selective for forebrain neurons,European Journal of Pharmacology, 1997, 331:117-126). Forebrains ofadult male CD rats are homogenized in 0.32M sucrose at 4° C. The crudenuclear pellet is removed by centrifugation at 1,000×g for 10 min., andthe supernatant centrifuged at 17,000×g for 25 min. The resulting pelletis resuspended in 5 mM Tris acetate pH 7.4 at 4° C. for 10 min. to lysecellular particles and again centrifuged at 17,000×g. The resultingpellet is washed twice in Tris acetate, resuspended at 10 mg protein/mland stored at −20° C. until use.

For binding assays, membranes are thawed, homogenized, and diluted to0.5 mg protein/ml with 50 mM Tris HCl, pH 7.4. Compounds under study areadded at various concentrations followed by racemic [³H] CP-101,606(specific activity 42.8 Ci/mmol, 5 nM final concentration). Followingincubation for 20 min at 30° C. in a shaking water bath, samples arefiltered onto Whatman GFB glass fiber filters using a MB-48R CellHarvester (Brandel Research and Development Laboratories, GaithersburgMd.). Filters are washed for 10 s with ice cold Tris HCl buffer and theradioactivity trapped on the filter quantified by liquid scintillationspectroscopy. Nonspecific binding is determined in parallel incubationscontaining 100 μM racemic CP-101,606. Specific binding is defined astotal binding minus nonspecific binding.

In one embodiment of the present invention, an NR2B subunit selectiveNMDA antagonist is furthermore selective for NR2B subunit-containingNMDA receptors over α₁-adrengergic receptors. For example, althoughifenprodil (supra) has selectivity for the NR2B subtype of NMDAreceptor, this compound is also a well known α₁-adrenergic receptorantagonist. (Carter et al. J. Pharmacol. Exp. Ther., 235, 475-482(1990)). Compounds that antagonize α₁-adrengergic receptors can cause areduction in blood pressure that can be a complication to therapeuticuse. Preferably, the NMDA antagonist has a ratio of NR2B receptoractivity to α₁-adrenergic receptor activity of at least about 3:1, morepreferably at least about 5:1.

Affinity for the NR2B subunit containing NMDA receptor is measured asthe IC₅₀ for displacement of specific binding of racemic[³H](+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanolfrom rat forebrain membranes (described above). Affinity for theα₁-adrengergic receptor is defined as the IC₅₀ for displacement ofspecific binding of racemic [³H]prazosin from rat brain membranes,measured as described by Greengrass and Bremner (Binding Characteristicsof [ ³H]prazosin to Rat Brain α-Adrenergic Receptors, European Journalof Pharmacology, 55, 323-326, (1979)). A compound with a ratio of([³H]prazosin/[³H](+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol)affinity greater than three is considered selective.

Forebrains of adult male Sprague Dawley rats are homogenized in 20volumes of ice cold 50 mM Tris/HCl buffer (pH 7.7). The homogenate iscentrifuged at 50,000×g for 10 min. at 4° C. The pellet is resuspendedand centrifuged under identical conditions and the final pellet isresuspended in 80 volumes of 50 mM Tris/HCl (pH 8.0) at 4° C.

For binding assays, compounds under study are added at variousconcentrations to 500 μg membrane protein in 1 ml of 50 mM Tris/HClbuffer, followed by [³H]prazosin (Amersham, specific activity 33Ci/mmol, 0.2 nM final concentration). Following incubation for 30 min at25° C. in a shaking water bath, samples are filtered onto Whatman GFBglass fiber filters using a MB-48R Cell Harvester (Brandel Research andDevelopment Laboratories, Gaithersburg Md.). Filters are washed threetimes for 10 s with ice cold Tris HCl buffer and the radioactivitytrapped on the filter quantified by liquid scintillation spectroscopy.Nonspecific binding is determined in parallel incubations containing100nM prazosin. Specific binding is defined as total binding minusnonspecific binding.

An effective amount of the NR2B subunit selective NMDA antagonist foruse on the present invention is typically from about 0.02 to 250mg/kg/day (0.001-12.5 g/day in a typical human weighing 50 kg) in singleor divided doses, regardless of route of administration. A morepreferred dosage range is from about 0.15 mg/kg/day to about 250mg/kg/day.

Of course, depending on the specific circumstances (for example, thepresence or absence of a predisposition to the disease or conditionbeing treated, the severity or expected severity of the disease, or theage or general health of the patient), even doses outside theaforementioned ranges may be in order. The particular dose given thespecific circumstances can be determined by a physician or otherhealth-care professional of ordinary skill.

The NR2B subunit selective NMDA receptor antagonist useful in the methodof the present invention is generally administered in the form of apharmaceutical composition comprising one or more NR2B subunit selectiveNMDA receptor antagonists together with a pharmaceutically acceptablecarrier or diluent.

The compositions described herein useful in the present invention aregenerally formulated in a conventional manner utilizing solid or liquidvehicles or diluents as appropriate to the mode of administration. Forpurposes of oral administration, tablets containing excipients such assodium citrate, calcium carbonate and dicalcium phosphate may beemployed along with various disintegrants such as starch and preferablypotato or tapioca starch, alginic acid and certain complex silicates,together with binding agents such as polyvinylpyrrolidone, sucrose,gelatin and acacia. Additionally, lubricating agents such as, but notlimited to, magnesium stearate, sodium lauryl sulfate and talc are oftenvery useful for tableting purposes. Solid compositions of a similar typemay also be employed as fillers in soft elastic and hard-filled gelatincapsules; preferred materials in this connection also include, by way ofexample and not of limitation, lactose or milk sugar as well as highmolecular weight polyethylene glycols. When aqueous suspensions and/orelixirs are desired for oral administration, the essential activeingredient may be combined with various sweetening or flavoring agents,coloring matter or dyes and, if so desired, emulsifying and/orsuspending agents, together with diluents such as water, ethanol,propylene glycol, glycerin and various like combinations thereof.

The present invention is illustrated by the following examples, but isnot limited to the details thereof.

All nonaqueous reactions were run under nitrogen for convenience andgenerally to maximize yields. All solvents/diluents were dried accordingto standard published procedures or purchased in a predried form. Allreactions were stirred either magnetically or mechanically. NMR spectraare recorded at 300 MHz and are reported in ppm. The NMR solvent wasCDCl₃ unless otherwise specified. IR spectra are reported in cm¹,generally specifying only strong signals.

EXAMPLE 1 Enantiomeric (1S,2S)- and(1R,2R)-1-(4-Hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-1-propanol

(+)-Tartaric acid (300 mg, 2 mmol) was dissolved in 30 mL warm methanol.Racemic1S*,2S*-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-1-propanol(655 mg, 2 mmol) was added all at once. With stirring and gentle warminga colorless homogeneous solution was obtained. Upon standing at ambienttemperature 24 hours, 319 mg (66%) of a fluffy white precipitate wasobtained. This product was recrystallized from methanol to give 263 mgof the (+)-tartrate salt of levorotatory title product as a white solid;mp 206.5-207.5° C.; [alpha]_(D)=36.2°. This salt (115 mg) was added to50 mL of saturated NaHCO₃. Ethyl acetate (5 mL) was added and themixture was vigorously stirred 30 minutes. The aqueous phase wasrepeatedly extracted with ethyl acetate. The organic layers werecombined and washed with brine, dried over calcium sulfate, andconcentrated. The tan residue was recrystallized from ethylacetate-hexane to give 32 mg (39%) of white, levorotatory title product;mp 203-204° C.; [alpha]_(D)=−58.4°. Anal. Calc'd. for C₂₀H₂₅NO₃; C,73.37; H, 7.70; N, 4.28. Found: C, 72.61; H, 7.45; N, 4.21.

The filtrate from the (+)-tartrate salt preparation above was treatedwith 100 mL saturated aqueous NaHCO₃ and extracted well with ethylacetate. The combined organic extracts were washed with brine, driedover calcium sulfate and concentrated to give 380 mg of recoveredstarting material (partially resolved). This material was treated with(−)-tartaric acid (174 mg) in 30 mL of methanol as above. After standingfor 24 hours, filtration gave 320 mg (66%) of product which was furtherrecrystallized from methanol to produce 239 mg of the (−)-tartrate saltof dextrorotatory title product; mp 206.5-207.5° C. [alpha]_(D)=+33.9°.The latter was converted to dextrorotatory title product in the mannerabove in 49% yield; mp 204-205° C.; [alpha]_(D)=+56.9°. Anal. Found: C,72.94; H, 7.64; N, 4.24.

EXAMPLE 2(1S,2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidin-yl)-1-propanolmethanesulfonate trihydrate

A 50 gallon glass lined reactor was charged with 17.1 gallons ofacetone, 8.65 kilograms (kg) (57.7 mol) of 4′-hydroxypropiophenone, 9.95kg (72.0 mol) of potassium carbonate and 6.8 liters (l) (57.7 mol) ofbenzylbromide. The mixture was heated to reflux (56° C.) for 20 hours.Analysis of thin layer chromatography (TLC) revealed that the reactionwas essentially complete. The suspension was atmosphericallyconcentrated to a volume of 10 gallons and 17.1 gallons of water werecharged. The suspension was granulated at 25° C. for 1 hour. The productwas filtered on a 30″ Lapp and washed with 4.6 gallons of water followedby a mixture of 6.9 gallons of hexane and 2.3 gallons of isopropanol.After vacuum drying at 45° C., this yielded 13.35 kg (96.4%) of theabove-depicted product.

A second run was carried out with 9.8 kg (65.25 mol) of4′-hydroxypropiophenone using the procedure described above. Afterdrying 15.1 kg (96.3%) of the above-depicted product was obtained.

Under a nitrogen atmosphere, a 100 gallon glass lined reactor wascharged with 75 gallons of methylene chloride and 28.2 kg (117.5 mol) ofthe product from step 1. The solution was stirred five minutes and then18.8 kg of bromine was charged. The reaction was stirred for 0.5 hoursat 22° C. Analysis of TLC revealed that the reaction was essentiallycomplete. To the solution was charged 37 gallons of water and themixture was stirred for 15 minutes. The methylene chloride was separatedand washed with 18.5 gallons of saturated aqueous sodium bicarbonate.The methylene chloride was separated, atmospherically concentrated to avolume of 40 gallons and 60 gallons of isopropanol was charged. Theconcentration was continued until a pot temperature of 80° C. and finalvolume of 40 gallons were obtained. The suspension was cooled to 20° C.and granulated for 18 hours. The product was filtered on a 30″ Lapp andwashed with 10 gallons of isopropanol. After vacuum drying at 45° C.,this yielded 29.1 kg (77.6%) of the above-depicted product.

Under a nitrogen atmosphere, a 20 gallon glass lined reactor was chargedwith 4.90 kg (15.3 mol) of the product from step 2, 7.0 gallons of ethylacetate, 2.70 kg (15.3 mol) of 4-hydroxy-4-phenylpiperidine and 1.54 kgof triethylamine (15.3 mol). The solution was heated to reflux (77° C.)for 18 hours. The resulting suspension was cooled to 20° C. Analysis byTLC revealed that the reaction was essentially complete. The byproduct(triethylamine hydrobromide salt) was filtered on a 30″ Lapp and washedwith 4 gallons of ethyl acetate. The filtrate was concentrated undervacuum to a volume of 17 liters. The concentrate was charged to 48liters of hexane and the resulting suspension granulated for 2 hours at20° C. The product was filtered on a 30″ Lapp and washed with 4 gallonsof hexane. After vacuum drying at 50° C., this yielded 4.9 kg (77%) ofthe above-depicted product.

A second run was carried out with 3.6 kg (11.3 mol) of the product fromstep 2 using the procedure described above. After drying 4.1 kg (87%) ofthe above-depicted product was obtained.

Under a nitrogen atmosphere, a 100 gallon glass lined reactor wascharged with 87.0 gallons of 2B ethanol and 1.7 kg (45.2 mol) of sodiumborohydride. The resulting solution was stirred at 25° C. and 9.4 kg(22.6 mol) of the product from step 3 was charged. The suspension wasstirred for 18 hours at 25-30° C. Analysis by TLC revealed that thereaction was essentially complete to the desired threo diastereoisomer.To the suspension was charged 7.8 liters of water. The suspension wasconcentrated under vacuum to a volume of 40 gallons. After granulatingfor 1 hour, the product was filtered on a 30″ Lapp and washed with 2gallons of 2B ethanol. The wet product, 9.4 gallons of 2B-ethanol and8.7 gallons of water were charged to a 100 gallon glass lined reactor.The suspension was stirred at reflux (78° C.) for 16 hours. Thesuspension was cooled to 250° C., filtered on 30″ Lapp and washed with 7gallons of water followed by 4 gallons of 2B ethanol. After air dryingat 50° C., this yielded 8.2 kg (86.5%) of the above-depicted product.This material was recrystallized in the following manner.

A 100 gallon glass lined reactor was charged with 7.9 kg (18.9 mol) ofthe product from step 3, 20 gallons of 2B ethanol and 4 gallons ofacetone. The suspension was heated to 70° C. producing a solution. Thesolution was concentrated atmospherically to a volume of 15 gallons. Thesuspension was cooled to 25° C. and granulated for 1 hour. The productwas filtered on a 30″ Lapp. The wet product and 11.7 gallons of 2Bethanol was charged to a 100 gallon glass lined reactor. The suspensionwas heated to reflux (78° C.) for 18 hours. The suspension was cooled to25° C., filtered on a 30″ Lapp and washed with 2 gallons of 2B ethanol.After air drying at 50° C. this yielded 5.6 kg (70.6%) of theabove-depicted product.

Under a nitrogen atmosphere, a 50 gallon glass lined reactor was chargedwith 825 g of 10% palladium on carbon (50% water wet), 5.5 kg (13.2 mol)of the product from step 4 and 15.5 gallons of tetrahydrofuran (THF).The mixture was hydrogenated between 40-50° C. for 2 hours. At thistime, analysis by TLC revealed that the reduction was essentiallycomplete. The reaction was filtered through a 14″ sparkler precoatedwith Celite and washed with 8 gallons of THF. The filtrate wastransferred to a clean 100 gallon glass lined reactor, vacuumconcentrated to a volume of 7 gallons and 21 gallons of ethyl acetatewere charged. The suspension was atmospherically concentrated to avolume of 10 gallons and a pot temperature of 72° C. The suspension wascooled to 10° C., filtered on a 30″ Lapp and washed with 2 gallons ofethyl acetate. After air drying at 55° C. this yielded a 3.9 kg (90%) ofthe above-depicted product (i.e., the free base).

A 100 gallon glass lined reactor was charged with 20 gallons of methanoland 3.7 kg (11.4 mol) of the product from step 5 (i.e., the free base).The suspension was heated to 60° C. and 1.7 kg (11.4 mol) ofD-(−)-tartaric acid were charged. The resulting solution was heated toreflux (65° C.) for 3 hours after which a suspension formed. Thesuspension was cooled to 35° C., filtered on a 30″ Lapp and washed with1 gallon of methanol. The wet solids were charged to a 100 gallon glasslined reactor with 10 gallons of methanol. The suspension was stirredfor 18 hours at 25° C. The suspension was filtered on a 30″ Lapp andwashed with 2 gallons of methanol. After air drying at 50° C. thisyielded 2.7 kg (101%) of the above-depicted product (i.e., the tartaricacid salt of the free base (R-(+)-enantiomer)). This material waspurified in the following manner:

A 100 gallon glass lined reactor was charged with 10.6 gallons ofmethanol and 2.67 kg (5.6 mol) of the above tartaric acid salt. Thesuspension was heated to reflux (80° C.) for 18 hours. The suspensionwas cooled to 30° C., filtered on a 30″ Lapp and washed with 4 gallonsof methanol. After air drying at 50° C., this yielded 2.05 kg (76.7%) ofthe above-depicted product (i.e., the tartaric acid salt of the freebase).

A 55 liter nalgene tub was charged with 30 liters of water and 1056 g(12.6 mol) of sodium bicarbonate at 20° C. To the resulting solution wascharged 2.0 kg (4.2 mol) of the product from step 6 (i.e., the tartaricacid salt of the free base). The suspension was stirred for 4 hoursduring which a great deal foaming occurred. After the foaming ceased,the suspension was filtered on a 32 cm funnel and washed with 1 gallonof water. After air drying at 50° C., this yielded 1.28 kg (93.5%) ofthe above-depicted product (i.e., the free base).

A 22 liter flask was charged with 1277 g (3.9 mol) of product from step7 and 14 liters of water. The suspension was warmed to 30° C. and 375 g(3.9 mol) of methane sulfonic acid were charged. The resulting solutionwas warmed to 60° C., clarified by filtering through diatomaceous earth(Celite™) and washed with 2 liters of water. The speck-free filtrate wasconcentrated under vacuum to a volume of 6 liters. The suspension wascooled to 0-5° C. and granulated for 1 hour. The product was filtered onan 18″ filter funnel and washed with 635 ml of speck-free water. Afterair drying at 25° C. for 18 hours, this yielded 1646 g (88%) of theabove-depicted product (i.e., the mesylate salt trihydrate).

EXAMPLE 3(1R*,2R*)-1-(4-hydroxy-3-methylphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol-mesylate

A mixture of 3-methyl-4-triisopropylsilyloxy-Ε-bromopropiophenone (9.17g, 22.97 mmol), 4-(4-fluorophenyl)-4-hydroxypiperidine (6.73 g, 34.45mmol) and triethylamine (8.0 mL, 57.43 mmol) in in ethanol (180 mL) wasrefluxed for 6 hours. The solvent was removed at reduced pressure andthe residue was partitioned between ethyl acetate and water. The phaseswere separated and the organic layer was washed with brine, dried overcalcium sulfate and concentrated. The residue was flash chromatographedon silica gel (3×3.5 inches packed in hexane) with elution proceeding asfollows: 10% ethyl acetate/hexane (1000 mL), nil; 20% ethylacetate/hexane (700 mL), nil; 20% ethyl acetate/hexane (1300 mL) and 25%ethyl acetate/hexane (600 mL), 7.66 g (65%) of1-(3-methyl-4-triisopropylsilyloxyphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-oneas a yellow foam which was suitable for use without furtherpurification. A sample recrystallization from ethyl acetate/hexane aswhite crystals had: m.p. 78-82° C.

A mixture of sodium borohydride (0.564 g, 14.92 mmol) and ethanol (60mL) was stirred 10 minutes and then1-(3-methyl-4-triisopropylsilyloxyphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-one(7.66 g, 14.92 mmol in 10 mL of ethanol) was added with two 30 mLethanol rinses. The reaction mixture was stirred at ambient temperatureovernight. The white solid that precipitated was collected by filtrationand dried to yield 5.72 g (74%) of(1R*,2R*)-1-(3-methyl-4-triisopropylsilyloxyphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol,which was suitable for use without further purification and had: m.p.188-89° C.

The product of the above reaction (5.72 g, 11.1 mmol) was dissolved intetrahydrofuran (150 mL) and tetrabutylammonium fluoride (12.21 mL,12.21 mmol, 1M tetrahydrofuran solution) was added. The reaction wasstirred 1 hour at ambient temperature and then concentrated. The residuewas partitioned between ethyl acetate and water and the two phases wereseparated. The organic layer was slurried with methylene chloride. Thewhite solid that precipitated was collected by filtration and dried toafford 3.41 g (85%) of(1R*,2R*)-1-(4-hydroxy-3-methylphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol.A sample (0.16 g, 0.447 mmol) was converted to the correspondingmesylate salt. The salt was slurried in methanol (8 mL) andmethanesulfonic acid (0.029 mL, 0.45 mmol) was added. The mixture wasfiltered and concentrated. The mixture was then recrystallized fromethanol to give 0.152 g (58%) of the mesylate salt which had: m.p.215-216° C. Analysis calculated for C₂₁H₂₅FNO₃.CH₄SO₃: C, 58.01; H,6.64, N, 3.07. Found: C, 57.99; H, 6.72; N, 3.17.

EXAMPLE 4 1R,2R1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenyl-piperidin-1-yl)-propan-1-oland 1S,2S1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenyl-piperidin-1-yl)-propan-1-ol

A mixture of 2-bromo-1-(2,2-diphenyl-benzo(1,3)dioxol-5-yl)-propan-1-one(2.00 g, 4.89 mmol), 4-hydroxy-4-phenylpiperidine (0.9 g, 5.08 mmol) andtriethylamine (1.40 mL, 10.04 mmol) in ethanol (50 mL) was refluxedovernight. The solvent was removed at reduced pressure and the residuewas partitioned between ether and water. The phases were separated andthe organic layer was washed with brine, dried over magnesium sulfateand concentrated. The residue was flash chromatographed on silica gel(2×5 inches packed with hexane) with elution proceeding as follows: 20%ethyl acetate/hexane (500 mL), unweighed forerun; 50% ethylacetate/hexane (500 mL), 1.76 g (71%) of1-(2,2)-diphenyl-benzo(1,3)dioxol-5-yl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-propan-1-oneas light tan foam which was suitable for use without furtherpurification and had: NMR δ 7.81 (dd, J=1.7, 8.3 Hz, 1H), 7.70 (d, J=1.6Hz, 1H), 7.64-7.13 (m, 15H), 6.92 (d, J=8.2 Hz, 1H), 4.07 (q, J=7.0 Hz,1H), 3.39-3.27 (m, 1H), 2.94-2.59 (m, #H), 2.30-2.04 (m, 2H), 1.74 (brt, J=13.2 Hz, 2H), 1.30 (d, J=6.8 Hz, 3H).

A mixture of sodium borohydride (0.15 g, 3.97 mmol) and ethanol (5 mL)was stirred 10 minutes and then1-(2,2-diphenyl-benzo(1,3)dioxol-5-yl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-propan-1-one (1.70 g, 3.36 mmol in 20 mL ofethanol) was added. The reaction was stirred at ambient temperature overthe weekend. The white precipitate was collected, rinsed with ethanoland ether and air dried to afford 1.35 g of crude product. The productwas recrystallized from ethanol/ethyl acetate/methylene chloride to give1.05 g (61%) of1R*,2R*)-1-(2,2-diphenyl-benzo(1,3)dioxol-5-yl)-2-(4-hydroxy-4-phenylpiperidin-1-yl)-propan-1-olwhich had: mp 224-224.5° C. Analysis calculated for C₃₃H₃₃NO₄: C, 78.08;H, 6.55; N, 2.76. Found: C, 78.16; H, 6.46N, 2.72.

A mixture of the product of the above reaction (1.00 g, 1.97 mmol) and10% palladium on carbon (0.175 g) in methanol (50 mL) and acetic acid(1.0 mL) was hydrogenated at 50 psi (initial pressure) for 5 hours atambient temperature. Additional catalyst (0.18 g) was added and thehydrogenation was continued overnight. The reaction was filtered throughdiatomaceous earth and the filter pad was rinsed with methanol. Thefiltrate was concentrated and the residue was partitioned between ethylacetate and saturated aqueous bicarbonate and stirred vigorously for 1hour. The phases were separated and the aqueous layer was extracted withethyl acetate (2×). The combined organic layer was washed with water andbrine, dried over magnesium sulfate and concentrated. The residue wasflash chromatographed on silica gel (1×4 inches) with elution proceedingas follows: 20% ethyl acetate/hexane (500 mL), nil; 10% methanol/ethylacetate (250 mL), 20% methanol/ethyl acetate (250 mL), and 50%methanol/ethyl acetate, 0.51 g (75%) of a light yellow-green solid. Thesolid was recrystallized from ethanol to afford(1R*,2R*)-1-(3,4-dihydroxyphenyl)-2-(4-hydroxy-4-phenyl-piperidin-1-yl)-propan-1-olas a white solid which had: mp 167-168° C. Analysis calculated forC₂₀H₂₅NO₄.0.5 C₂H₆O: C, 68.83; H, 7.70; N, 3.82. Found: C, 68.78; H,8.05; N, 3.70.

The racemic product was dissolved in ethanol and separated intoenantiomers by HPLC using the following chromatographic conditions:Column, Chiralcel OD; mobile phase, 25% ethanol/75% hexane; temperature,ambient (approximately 22° C.); detection, UV at 215 nM. Under theseconditions, 1R,2R1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenyl-piperidin-1-yl)propan-1-ol eluted with a retention time of approximately 9.12 minuntesand 1S,2S1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenyl-piperidin-1-yl)propan-1-ol eluted with a retention time of approximately 16.26 minutes.

EXAMPLE 5(3R*,4S*)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-chroman-4,7-diol

A mixture of 7-benzyloxy-3,3-dibromochromanone (54.7 g, 133 mmol),4-(4-fluorophenyl)-4-hydroxypiperidine (52.0 g, 266 mmol), andtriethylamine (38 mL, 270 mmol) in acetonitrile (2.5 L) was stirred 16hours at ambient temperature. A yellow precipitate formed and wascollected, washed well with water and ether, and air dried. The yield of7-benzyloxy-3-{4-(4-fluorophenyl)-4-hydroxy-piperidine-1-yl]-chromenonewas 55.4 g (93%) which was suitable for use without furtherpurification. A sample recrystallized from ethanol/tetrahydrofuran hadmp 220-221° C.: NMR DMSO_(∂σ)δ7.99 (d, J=9 Hz, 2H), 7.56-7.40 (m, 8H),7.18-7.08 (m, 4H), 5.25 (s, 2H), 5.06 (s, 1H), 3.60(br s, 1H), 3.55-3.35(m, 1H, partially obscured by water from the NMR solvent), 3.10-2.95 (m,2H), 2.15-2.00 (m, 2H), 1.71 (br t, J=13.7 Hz, 2H).

Analysis calculated for C₂₇H₂₄FNO₄: C, 72.80; H, 5.43; N, 3.13. Found:C, 72.83; H, 5.82; N, 2.82.

To a slurry of7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidine-1-yl]-chromenone(8.24 g, 18.5 mmol) in ethanol (400 mL) and tetrahydrofuran (600 mL) wasadded sodium borohydride (7.0 g, 185 mmol). The mixture was stirredovernight. Additional sodium borohydride (7.0 g) was added and thereaction mixture was stirred for 3 days. Water was added and the solventwas removed at reduced pressure at 45° C. The solids which formed werecollected and washed well with water and then ether. The solid wasfurther dried in vacuo overnight to give 5.01 g, 60% of3R*4S*7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4-olwhich was suitable for use without further purification. A samplerecrystallized from ethyl acetate/chloroform had mp. 194-195° C.; NMRδ7.56-7.30 (m, 8H), 7.06 (long range coupled t, J=8.7 Hz, 2H) 6.63 (dd,J=2.4, 8.5 Hz, 1H), 6.47 (d, J=2.4 Hz, 1H), 5.04 (s, 2H), 4.77 (d,J=4.5Hz, 1H), 4.37 (dd, J=3.5, 10.4Hz, 1H), 4.13 (t, J=10.4 Hz, 1H),3.82 (brs, 1H), 3.11 (br d, J=11.2 Hz, 1H), 2.92-2.71 (m, 4H), 2.21-2.06(m, 2H), 1.87-1.73 (m, 2H), 1.54 (s,1H).

Analysis calculated for C₂₇H₂₈FNO₄: C, 72.14; H, 6.28; N, 3.12. Found C,72.15; H, 6.21; N, 3.12.

A mixture of3R*4S*7-benzyloxy-3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4-ol(0.80 g, 1.78 mmol), 10% palladium on carbon (0.16 g), methanol (40 mL),and acetic acid (0.8 mL) was hydrogenated for 8 hours with a startingpressure of 48.5 psi. The reaction was filtered through celite and thefiltrate was concentrated. The residue was stirred vigorously with etherand saturaturated sodium bicarbonate for 1 hour. The solid was washedwith water and ether and dried in vacuo. Recrystallization from ethanolyielded 0.35 g (54%) of3R*4S*3-[4-(4-fluorophenyl)-4-hydroxy-piperidin-1-yl]-chroman-4,7-diolas a white solid which had mp 159-160° C.; NMR DMSO_(∂σ)δ7.55-7.47 (m,2H), 7.11 (t, J=9 Hz, 2H), 7.02 (d, J=8.4 Hz, 1H)k, 6.32 (dd, J=2.3, 8.3Hz, 1H), 6.15 (d, J=2.3 Hz 1H), 5.10-4.50 (br m with s at 4.63, 3H),4.23 (dd, J=2.8, 10.3 Hz, 1H), 4.04 (t, J=10.5 Hz, 1H), 2.99 (br d,J=10.8 Hz, 1H), 2.86 (br d, J=10.7 Hz, 1H), 2.73-2.50 (m, 3H), 2.08-1.90(m, 2H), 1.58 (br d, J=13 Hz, 2H).

Analysis calculated for C₂H₂₂FNO₄.0.25H₂O; C, 66.01; H, 6.23; N, 3.85.Found: C, 66.22; H, 6.58; N. 3.46.

1. A method of treating Restless Leg Syndrome in a mammal, wherein themethod comprises administering to the mammal an amount of an NR2Bsubunit selective NMDA antagonist, wherein the amount is effective intreating Restless Leg Syndrome, wherein the NR2B subunit selective NMDAreceptor antagonist is a compound of the formula

or a pharmaceutically acceptable acid addition salt thereof, wherein:(a) R² and R⁵ are taken separately and R¹, R², R³ and R⁴ are eachindependently hydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷ and R₅ ismethyl or ethyl; or, (b) R² and R⁵ are taken together and are

 forming a chroman-4-ol ring, and R¹, R³ and R⁴ are each independentlyhydrogen, (C₁-C₆) alkyl, halo, CF₃, OH or OR⁷; R⁶ is

R⁷ is methyl, ethyl, isopropyl or n-propyl; R⁸ is phenyl optionallysubstituted with up to three substituents independently selected fromthe group consisting of (C₁-C₆) alkyl, halo and CF₃; X is 0, S or(CH₂)_(n); and n is 0, 1, 2, or
 3. 2. A method according to claim 1,wherein the NR2B subunit selective NMDA receptor antagonist is(+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propanol;(1S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(-4-hydroxy-4-phenylpiperidino)-1-propanol;(3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-chroman-4,7-diol;a pharmaceutically-acceptable acid addition salt of one of saidcompounds; or(1R*,2R*)-1-(4-hydroxy-3-methylphenyl)-2-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol-mesylate.